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PubMed1995

The BM88 antigen, a novel neuron-specific molecule, enhances the differentiation of mouse neuroblastoma cells.

The BM88 antigen is a neuron-specific molecule widely distributed in the mammalian nervous system. It is a 22-kDa, apparently not glycosylated, integral membrane protein, which appears early during brain development and remains at high levels in the mature animal. Here, we describe the cDNA cloning of the porcine BM88 antigen and present evidence that this protein is involved in neuroblastoma cell differentiation. The deduced protein is a novel molecule consisting of 140 amino acids and bears a putative transmembrane domain at the COOH-terminal region. The mRNA of this protein is expressed only in neural tissues, where it is restricted to neurons. Stably transfected Neuro-2a cells overexpressing the BM88 antigen exhibited a significant change in morphology, reflected by enhanced process outgrowth, and a slower rate of division. Moreover, in the presence of differentiation agents, such as sucrose and retinoic acid, an accelerated differentiation of the transfected Neuro-2a cells was observed. Especially in the presence of sucrose, the consequent overexpression of the BM88 antigen in the transfected cells resulted in their enhanced morphological differentiation accompanied by the induction of neurofilament protein expression. Our results suggest that the BM88 antigen plays a role in the differentiation of neuroblastoma cells.

Amino Acid Sequence↗
PubMed1996

A tau promoter region without neuronal specificity.

The microtubule-associated protein tau is produced from a 6-kb mRNA expressed primarily in neurons. A 2-kb tau mRNA has also been characterized, which produces a tau isoform that localizes to the nucleus, and an 8-kb mRNA is expressed in the PNS. Mapping and sequencing of the human tau gene start showed that it has an unusually GC-rich 5'-untranslated region coded by a single untranslated exon (designated -1). Primer extensions and expression assays indicated that upstream of exon -1 is a promoter that is not neuron specific. This region contains consensus binding sites for transcription factors AP2, Sp1, and GCF.

Base Sequence↗
PubMed1984

Large scale preparation and crystallization of neuron-specific enolase.

A simple method has been developed for the large scale purification of neuron-specific enolase [EC 4.2.1.11]. The method consists of ammonium sulfate fractionation of brain extract, and two subsequent column chromatography steps on DEAE Sephadex A-50. The chromatography was performed on a short (25 cm height) and thick (8.5 cm inside diameter) column unit that was specially devised for the large scale preparation. The purified enolase was crystallized in 0.05 M imidazole-HCl buffer containing 1.6 M ammonium sulfate (pH 6.39), with a yield of 0.9 g/kg of bovine brain tissue.

Amino Acids↗
PubMed1986

A rapid purification of synapsin I: a neuron specific spectrin binding protein.

We have developed a one chromatographic step isolation protocol for the neuron specific protein synapsin I. This procedure results in a yield of 80 micrograms/g brain, which is ten fold better than the highest yield yet reported for this protein. The authenticity of the synapsin I isolated by this procedure is demonstrated by comigration with authentic synapsin I on SDS-polyacrylamide gels, crossreactivity with antibody specific against synapsin I, and nearly identical two dimensional chrymotryptic iodopeptide maps of authentic synapsin I and the protein purified by this protocol. Synapsin I isolated by this procedure retains its functional properties, demonstrated by the ability of synapsin I to stimulate the formation of a brain spectrin(240/235)/synapsin I/F-actin ternary complex as determined by a low shear falling ball viscometry assay. This novel protocol therefore has the advantage of being a rapid, high yield procedure that retains the functional properties of synapsin I.

Actins↗
PubMed1983

Neurone-specific enolase and creatine phosphokinase are protein components of rat brain synaptic plasma membranes.

Neuron-specific enolase and creatine phosphokinase were found, by 2-dimensional gel analysis, in rat brain synaptic plasma membranes (SPM). The identity of these enzymes was confirmed by comigration with purified rat brain NSE and CPK and by peptide analysis. The specific enzymatic activities of enolase and creatine phosphokinase, as well as of pyruvate kinase, also present on the membranes, were comparable to those in the homogenates when these three enzymes were fully activated. In the SPM all three enzymes, particularly enolase, were partially cryptic in that enzymatic activities were very low unless the membranes were treated with Triton X-100. They were resistant to both low-salt and high-salt extraction and to trypsin, except when Triton X-100 was present. These results suggest that the enzymes are tightly bound protein components of the membrane and that they may constitute an assembly capable of generating ATP.

Animals↗
PubMed1985

Neuron-specific enolase-like immunoreactivity in inner hair cells but not outer hair cells in the guinea pig organ of Corti.

Neuron-specific enolase (NSE) has been localized only in neurons and cells with characteristics of neurons. The immunocytochemical localization of NSE was examined in guinea pig cochleae to determine if hair cells, which have some neuronal characteristics, would show NSE-like immunoreactive labeling. NSE-like immunoreactivity was seen in inner hair cells but not in outer hair cells. This is the first report of NSE-like immunoreactivity in a receptor cell. NSE-like immunoreactivity was also seen in efferent fibers and terminals and in both type I and type II spiral ganglion cells. The finding of NSE-like immunoreactivity in inner but not outer cells adds to the number of differences found between them and may be related to differences in function and action.

Animals↗
PubMed1993

Detection of neuron-specific gamma-enolase messenger ribonucleic acid in normal human leukocytes by polymerase chain reaction amplification with nested primers.

BACKGROUND: NNE (non-neuronal alpha-enolase) is a glycolytic enzyme detected in most tissues. NSE (neuron-specific gamma-enolase) is detected in normal neurons and tumors such as neuroblastoma. Staining with antibodies against NSE is therefore used to detect neuroblastoma cells invading bone marrow. Since staining of normal leukocytes has been reported we asked whether bona fide NSE is in fact expressed in normal blood and marrow. EXPERIMENTAL DESIGN: We designed nested coding region specific primers for NSE and NNE and, after reverse transcription of mRNA, we amplified the coding region between these primers in a semi-nested polymerase chain reaction. In order to distinguish both iso-mRNAs from each other, we amplified a long (1,047 bp) template in a first round of 30 cycles with primers specific for NNE or NSE. One percent of this product was used in a second round of 30 cycles in which both sense primers and two nested anti-sense primers of alternate specificities yielding shorter products of discernible sizes (768 bp or 619 bp) were added together in the same reaction tube. With this combination of four primers, only that shorter product was amplified to visibility, the specificity of which was homologous to the template produced in the first 30 cycles. Restriction enzyme digestion of the amplified products was used to verify this polymerase chain reaction-based approach for the distinction of isoforms of RNA. RESULTS: This semi-nested polymerase chain reaction clearly allows for the distinction of mRNA for NNE or NSE and shows the presence of transcripts for NSE in normal human leukocytes from blood and bone marrow. CONCLUSIONS: This method exploiting short stretches of nucleotide differences in the coding regions for priming can more generally be applied to the distinction of all isoforms of RNA where nested specific primers can be designed. However, the presence of NSE specific transcripts in normal human leukocytes invalidates the use of this highly sensitive method as a disease marker in neuroblastoma.

Base Sequence↗
PubMed1995

Diagnostic significance of serum neuron-specific enolase and myelin basic protein assay in patients with acute head injury.

BACKGROUND: Neuron-specific enolase (NSE) and myelin basic protein (MBP) in the peripheral venous blood (PVB) have been reported to be sensitive markers for judging the prognosis of patients with head injury. However, to our knowledge, the levels of NSE and MBP in the internal jugular venous blood (IJVB) have never been studied. METHODS: In 25 patients with acute head injury, blood samples were taken from the internal jugular vein and the peripheral vein at the same time before any medical or surgical procedure was performed. The levels of NSE and MBP in the both venous blood samples were measured. The time interval between injury and sampling was 1.5-8.0 hours (mean 4.3 hours). The levels of NSE and MBP in the IJVB were compared to those in the PVB. The relationship between the clinical outcome and the serum levels of those was evaluated. RESULTS: The levels of NSE and MBP in the IJVB were almost equal to those in the PVB. The levels of NSE and MBP were significantly higher in the patients who died than in those who survived. In the survivors, the levels of NSE and MBP in the IJVB were 17.6 +/- 11.4 ng/ml and 1.4 +/- 1.5 ng/ml, whereas in the patients who died, both levels were elevated to 51.3 +/- 27.3 ng/ml (p < 0.005) and to 11.3 +/- 9.5 ng/ml (p < 0.01), respectively. CONCLUSIONS: The assay of serum NSE and MBP levels provides a reliable laboratory indicator of the degree of brain damage and allows early prediction of the prognosis in patients with acute head injury.

Acute Disease↗
PubMed1988

Levels of neuron-specific enolase after chemotherapy do not predict a response in small cell lung cancer.

Neuron-specific enolase (NSE) was measured in serum samples of 35 patients with small cell lung cancer and 10 control patients. The samples were collected during 10 days after the first course of chemotherapy, in order to investigate whether changes of NSE had a predictive value of tumour response. Three patterns of change of NSE were observed. Pattern 1 showed an increase of serum NSE with a maximum value more than 1.5 times the pretreatment level (n = 17); pattern 2 involved no increase at all or less than 1.5 times the pretreatment level (n = 14); pattern 3 showed a continuous decrease (n = 5). No relationship between the three patterns of change and the tumour response was observed. Only an NSE level less than 10 ng/ml at the time of start of the second course predicted a major response.

Carcinoma, Small Cell↗
PubMed1985

Immunohistochemical study of neuron specific enolase and S-100 protein in Hirschsprung's disease.

The distribution of whole differentiated neurons in the intestines from 15 children with Hirschsprung's disease was investigated using neuron specific enolase (NSE) and the perineuronal elements were studied using S-100 protein immunostaining. In aganglionic segments, NSE immunoreactive ganglion cells and S-100 positive satellite cells were absent, but the hypertrophic nerve trunks did show a markedly positive NSE and S-100 immunoreactivity. Two different forms of aganglionic segment were present. One was the middle aganglionic segment of long segment aganglionosis which was almost completely dennervated. In the other type, there were several NSE positive nerve fibers in the muscularis propria of both the aganglionic segment of short segment aganglionosis and the distal aganglionic segment of long segment aganglionosis. These latter two aganglionic segments seemed to be innervated by extrinsic nerves.

Adolescent↗
PubMed1985

Onset and development of neuron-specific enolase immunoreactivity in the peripheral vestibular system of the mouse.

The development of neuron-specific enolase (NSE) immunoreactivity in mouse sensory and ganglion vestibular cells was studied from gestation day 14 to adulthood. NSE staining appeared sequentially in these structures with a pattern that closely paralled their maturation sequences. The onset of NSE reactivity, at gestation day 15, in ganglion cells was concomitant with the formation of contacts between the afferent fibers and sensory cells, and was observed in hair cells, at gestation day 17, during the period in which the first synaptic structures form. The development of NSE staining in the cristae revealed an apex-base and axial gradients of maturation.

Animals↗
PubMed1999

Do blood levels of neuron-specific enolase and S-100 protein reflect cognitive dysfunction after coronary artery bypass?

BACKGROUND: Neuron-specific enolase (NSE) and S-100 protein have been used as markers for major brain damage. Cognitive dysfunction after cardiac surgery represents subtle brain damage that is detected by neuropsychological testing. We hypothesised that blood levels of NSE and S-100 protein would increase after coronary artery bypass surgery and that the change in blood level would correlate with the change in cognitive function. METHODS: We included 35 elderly patients undergoing coronary artery bypass surgery. Neuropsychological testing was performed before surgery and postoperatively at discharge from hospital and after 3 months using a neuropsychological test battery. Serum levels of NSE and S-100 protein were measured before surgery and postoperatively after 24 h and 48 h. RESULTS: Serum level of both NSE and S-100 protein increased significantly 24 and 48 h after coronary artery bypass. Only the increase in NSE after 24 h was found to correlate with the degree of cognitive dysfunction at discharge from hospital. The increase in S-100 protein after 24 h correlated significantly with the duration of cardiopulmonary bypass. CONCLUSION: NSE seems to be a useful blood marker for early cognitive dysfunction after coronary artery bypass surgery.

Aged↗
PubMed2004

Cerebrospinal fluid neuron-specific enolase in non-selected patients.

The degree to which cerebrospinal fluid (CSF) neuron-specific enolase (NSE) contributes to the diagnosis and prognosis of disorders of the central nervous system (CNS) or peripheral nervous system (PNS) is still under debate. The aim of the study was thus to assess the validity of CSF-NSE levels in the diagnostic work-up of these conditions. The study consecutively included 106 adult patients who had undergone a diagnostic spinal tap or myelography during the diagnostic work-up for various CNS or PNS disorders. Thirty-five of these patients (16 F, 19 M, aged 24-88 years) without indication of a CNS disorder and with normal routine CSF investigations served as controls. The remaining 71 patients (31 F, 40 M, aged 28-87 years) constituted the disease group. CSF-NSE was independent of sex and age. The upper reference limit of CSF-NSE was 0.01536 ng/L. CSF-NSE was elevated in 13 of the 71 patients (18%): 6 with metabolic myopathy, 4 with polyneuropathy and 3 with hepatic encephalopathy, multiple sclerosis and paraspasticity, respectively. Only 6 of the 13 patients (46%) showed CNS involvement. The study shows that CSF-NSE is elevated in only one-fifth of unselected patients who consecutively undergo a spinal tap. CSF-NSE is elevated most frequently in patients with metabolic myopathy and polyneuropathy, even in cases without CNS abnormalities.

Adult↗
PubMed2000

Do blood concentrations of neurone specific enolase and S-100 beta protein reflect cognitive dysfunction after abdominal surgery?ISPOCD Group.

Neurone specific enolase (NSE) and S-100 beta protein have been used as markers of brain damage. We hypothesized that blood concentrations of NSE and S-100 beta protein reflect cognitive dysfunction after abdominal surgery. We studied 65 elderly patients in whom neuropsychological testing was performed before abdominal surgery, at discharge from hospital and after 3 months. Serum concentrations of NSE and S-100 beta protein were measured before surgery and after 24, 48 and 72 h. Serum concentrations of S-100 beta protein increased significantly while NSE concentrations decreased significantly. The increase in S-100 beta protein concentration after 48 h was significantly greater in patients with delirium. No correlation was found between cognitive dysfunction and S-100 beta protein or NSE concentration. We conclude that blood concentrations of S-100 beta protein increase after abdominal surgery and may be related to postoperative delirium.

Abdomen↗
PubMed1990

Postnatal development of a granule cell-enriched, neurone-specific glycoprotein, gp50, in normal and thyroid-deficient rats.

Glycoprotein gp50 is a neurone-specific, granule cell-enriched glycoprotein that is also a major component of isolated synaptic membranes. Here, we describe the use of a monoclonal antibody, mab SM gp50, to study the postnatal development of gp50 in the brain of normal and thyroid-deficient rats. Radioimmunoassay, enzyme-linked immunosorbent assay, and Western blotting show that gp50 is not detectable in brain until postnatal day 4 (P4) in both forebrain and cerebellum. In forebrain, the rate of increase of gp50 levels is maximal between P12 and P20. It is somewhat later in cerebellum, where peak levels are attained between P30 and P35. Immunocytochemical studies show little detectable gp50-like immunoreactivity before P16, and the staining is still weak, relative to adult tissue, at P25. The intense staining of the granule cell layer characteristic of adult cerebellum predominantly appears after P25. Development of gp50 is severely retarded in the cerebellum of thyroid-deficient rats, particularly during the second and third postnatal weeks. However, by the fourth postnatal week, gp50 levels in normal and hypothyroid animals are comparable. The results indicate that significant alterations in the pattern of gp50 expression continue to occur at a late stage of cerebellar development. In particular, the increase in immunocytochemical staining of the granule cells after P25 is striking in that by this time most major events associated with cerebellar development are essentially complete.

Animals↗
PubMed1995

[Clinical value of measurement of serum creatine kinase BB and neuron specific enolase for small cell lung cancer].

Serum creatine kinase BB (CK-BB) and neuron specific enolase (NSE) were measured with chromatography-fluorometric method and ABC-ELISA in 20 patients with small cell lung cancer (SCLC), 30 patients with non small cell lung cancer (NSCLC), 25 patients with benign pulmonary diseases (BPD) and 30 healthy subjects (C). The results revealed that serum concentrations of CK-BB and NSE in SCLC were significantly greater than those in other three groups (P < 0.001). The mean values of CK-BB and NSE in SCLC, NSCLC, BPD, C were 30.2, 8.4, 6.3, 4.3 IU/L and 52.2, 12.7, 10.3, 9.2 ng/ml, respectively. If values above 9.5 IU/L (CK-BB) and 20.8 ng/ml (NSE) were considered abnormal, 70% and 80% of the values of CK-BB and NSE in SCLC were positive, respectively, which were higher than those in NSCLC (P < 0.005). The serum levels of CK-BB had linear correlation with those of NSE in SCLC (r = 0.7934, P < 0.001). By combined determination, the sensitivity and specificity were increased to 90% and 94% respectively. Therefore, the results suggest that serum CK-BB and NSE be diagnostic markers for SCLC. Concurrent determination may raise their value in clinical use.

Adult↗
PubMedDate not supplied

[Assessment of the informative value of neuron-specific enolase determined by the immunoenzyme method].

The diagnostic informative value of measurements of neuron-specific enolase (NSE) in health, pulmonary cancer (including small-cell one), 8 different diseases, and pregnant patients with gestation periods from 5 to 33 weeks was assessed. Comparison of the results in each of these 10 groups with the results in health controls did not reveal any appreciable differences for any of the groups or between the tested groups. The authors consider that NSE measurements cannot be recommended for use at other than cancer institutions.

Biomarkers, Tumor↗
PubMed1994

Serum neuron-specific enolase levels in patients with small cell carcinoma of the uterine cervix.

Serum neuron-specific enolase (NSE) is a tumor marker for neuroendocrine tumors or tumors of neuroepithelial origin. While the clinical application of NSE in small cell lung cancer has been established, its role in small cell carcinoma of the uterine cervix is unknown. We examined serum NSE in six patients with cervical small cell carcinoma and 13 patients with cervical squamous cell carcinoma. Elevated serum NSE was noted in four of six patients (67%) with small cell carcinoma, but none of the 13 patients with squamous cell carcinoma. An extremely high NSE serum level (154.7 ng/mL) was found in one patient with recurrence of small cell carcinoma, suggesting its correlation with disease extension. Although further studies to evaluate its relevance to prognosis, treatment effect, and early detection of recurrence are in progress, this preliminary result seems promising.

Adult↗